T26_26-3
DNA from up to 200 l of skin mucus samples was extracted using a High Pure PCR Template Preparation Kit (Roche, Basel, Switzerland), including a lysozyme lysis step for optimized DNA extraction (Piazzon et al., 2019).For the skin mucus samples, the complete 16S rRNA gene (V1-V9) was sequenced using the ONT MinION (Oxford Nanopore Technologies, Oxford, UK) device and the 16S Barcoding Kit 1-24 (SQK-16S024) v.16S_9086_v1_revR_14Aug2019, according to the manufacturers protocol including modifications of input DNA and PCR conditions described elsewhere (Toxqui-Rodrguez et al., 2022). The amplified DNA was quantified using PicoGreen (Life Technologies, Carlsbad, CA, USA), and libraries of 100 fmol were loaded into the ONT MinION device. Libraries were sequenced using an R9.4/FLO-MIN106 flow cell and demultiplexed using the MinKNOW v21.11.17. The sequencing was stopped when approximately an average of 100,000 reads per sample was achieved, which constituted a sequencing run time of 21-23 h.